The long-term objective is to investigate possible links between gluten ingestion, zonulin-dependent sustained increase in intestinal permeability, and the onset of Type 1 diabetes in order to develop strategies for the treatment of the disease, and eventually methods for prevention as well. Several studies suggest that an increased intestinal permeability due to alteration of intestinal tight junctions, mediated by zonulin, can be involved in the pathogenesis of autoimmune diseases, including Type 1 diabetes. Gluten has been implicated as one of the possible environmental triggers involved in type 1 diabetes pathogenesis. We have developed a novel mechanistic approach, based on identification of patients with alteration of the zonulin system, to evaluate the role of gluten in the autoimmune destruction of pancreatic beta cells. The primary specific aims of this proposal are to 1) evaluate zonulin and HLA type as a marker for the development of Type 1 diabetes, and 2) evaluate peripheral blood mononuclear cell (PBMC) cytokine expression in children with elevated zonulin and new onset Type 1 diabetes following a gluten-free diet (GFD). This diet intervention is being evaluated for its effect on the preservation of pancreatic beta-cell mass in children newly diagnosed with Type 1 diabetes. This strategy, if proven successful could preventing further loss of pancreatic beta cells. The first aim is addressed by assaying zonulin in samples collected serially in children at risk for the development of T1D. For the second aim, children with elevated zonulin will be randomized, 15 children with new onset diabetes in the control (normal diet) group, 15 children in the intervention (gluten-free diet) group, and evaluated serially for PBMC cytokines, pancreatic beta-cell reserve (by C-peptide stimulation testing) and zonulin at baseline, and then at 3 month intervals for one year. Relevance to public health: The goal of this study is to evaluate the role of zonulin as a marker for T1D risk, and the role of zonulin and cytokines as a marker for autoimmune progression in T1 D. If successful, these markers could be used for detecting at risk patients for targeting prevention strategies, and also in evaluating the success of interventions for children with new onset diabetes. [unreadable] [unreadable] [unreadable]